NIR-FLIVO 690 Tracer In vivo Assay
FLIVO® (FLuorescence in vIVO) is a powerful method for non-invasive detection of caspase activity in vivo. NIR-FLIVO 690 Tracers are non-cytotoxic, cell-permeant fluorescent inhibitors of caspases optimized for use in whole live animals. ICT’s NIR-FLIVO 690 Tracer probe contains the preferred binding sequence for most caspases (Val-Ala-Asp or VAD). This preferred poly caspase tripeptide binding sequence is labeled with *Dylight® 690 dye and a fluoromethyl ketone (FMK) reactive entity.
FLIVO® is a registered trademark of ICT. *Dylight® is a registered trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
NIR-FLIVO 690 Tracer In vivo Assay
FLIVO® kits provide a simple and accurate method to detect caspase activity in vivo. Like our FLICA® probes but optimized for whole live animal labeling. To label cells containing active caspases, inject FLIVO intravenously and let it circulate ~60 minutes. Because the reagent is cell-permeant, it readily diffuses in and out of all cells that it encounters as it circulates throughout the body. If there are active caspase enzymes inside a cell, FLIVO will form an irreversible covalent bond with the caspase active site. The bound FLIVO probe will remain inside the cell if the cell membrane is intact. Any unbound FLIVO is removed from the circulation of the animal in about an hour. The remaining near-infrared fluorescent signal in the tissue is a direct measure of caspase activity that occurred at the time the reagent was injected. Once the excess FLIVO has cleared from the body of the animal, the animal is ready for analysis. No further staining is necessary. Apoptotic cells and tissues can be viewed using whole live animal imagers. Alternatively, target tissues can be removed and processed for analysis. FLIVO is very sensitive and will pick up naturally occurring background apoptosis. Apoptotic cells have more active caspases than control cells, therefore they fluoresce brighter with FLIVO.
NIR-FLIVO® 690 Tracer (690-VAD-FMK)
Poly Caspases
690 nm/709 nm
Use instrumentation designed for non-invasive live animal imaging, alternatively, excised tissues can be analyzed by fluorescence microscope or flow cytometer.
Whole animal fluorescence imaging, excised tissues
-20°C
Ships overnight (domestic), International Priority Shipping
United States
- Prepare samples and controls.
- Dilute 10X Injection Buffer 1:10 with 45 mL diH20.
- Reconstitute NIR-FLIVO 690 Tracer with 50 μL DMSO.
- Dilute NIR-FLIVO 1:10 with 450 μL 1X Injection Buffer.
- Inject 50 μL intravenously.
- Let NIR-FLIVO circulate for 4 hours.
- Image the animal. NIR-FLIVO 690 Tracer excites at 690 nm and emits at 709 nm.
- If not viewing directly, excise tissue and process samples for further analysis.
- Optional: label excised samples with additional stains, such as Hoechst 33342, or an antibody.
- If desired, fix excised samples.
- Analyze excised samples with a fluorescence microscope or flow cytometer.
Kit 9112:
2 vials of NIR-FLIVO® 690 Tracer (*Dylight® 690-VAD-FMK), 10 Tests per vial, 71.4 μg per vial, #6306
1 bottle of 10X Injection Buffer, 5 mL, #6220
Kit Manual
FLIVO® is a registered trademark of ICT. *Dylight® is a registered trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.